GENES FOR AXON GUIDANCE IN THE DROSOPHILA VISUAL SYSTEM

果蝇视觉系统中轴突引导的基因

• 批准号: 6084015
• 负责人: Qi He
• 金额: $ 11.33万
• 依托单位: BROOKLYN COLLEGE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6084015
• 关键词: Drosophilidae; animal genetic material tag; axon; biomarker; cell differentiation; cell population study; confocal scanning microscopy; cytogenetics; gene expression; genetic library; molecular cloning; neuronal guidance; nucleic acid sequence; retina; synapses; tissue mosaicism; visual photoreceptor

DESCRIPTION (Adapted from applicant's abstract): The broad, long-term goal of this application is to investigate the cellular and molecular mechanisms involved in establishing the precise axon connections between retinal photoreceptor neurons and their target neurons in the brain of Drosophila. The specific aims are: (1) to use genetic mosaic analysis to determine which cell populations in the visual system require the function of a particular gene; (2) to use cell-type specific markers to determine which cell populations are present and differentiate normally in the mutant animals; (3) to characterize at the molecular level genes selected on the basis of the analyses described in aims 1 and 2. Mosaic analysis will be used to determine cell populations that require the function of a particular gene, and the generation of mosaic animals will use the yeast FLP recombinase/FLP system. Antibodies recognizing specific cell types will be used to determine cell identity in mutant animals. In addition, mutant strains with specific cell populations marked with a reporter gene lacZ will be created and different cell types determined by an anti-beta-galactosidase antibody. The analysis will include confocal microscopy. The genes identified in these analyses will be characterized by molecular cloning, where Drosophila cDNA libraries will be screened using probes from plasmid rescue, and full-length sequences of mutant genes obtained. Finally, phenotype-rescuing analysis through mRNA injection will be performed to confirm the rescuing ability of a cloned gene.

描述（改编自申请人的摘要）： 该应用是为了研究细胞和分子机制 参与了视网膜之间的精确轴突连接 果蝇大脑中的感光神经元及其靶神经元。这 具体目的是：（1）使用遗传镶嵌分析来确定哪个细胞 视觉系统中的种群需要特定基因的功能。 （2） 使用细胞类型的特定标记来确定哪些细胞群是 在突变动物中正常出现和区分； （3）表征 根据分子水平基因根据所述的分析选择 目标1和2。马赛克分析将用于确定细胞群体 需要特定基因的功能，并产生镶嵌动物 将使用酵母FLP重组酶/FLP系统。识别特异性的抗体 细胞类型将用于确定突变动物中的细胞身份。在 此外，具有记者标记的特定细胞群体的突变菌株 将创建Gene Lacz，并由不同的细胞类型确定 抗β-半乳糖苷酶抗体。分析将包括共焦 显微镜。这些分析中鉴定的基因将以以下特征 分子克隆，将使用果蝇cDNA文库进行筛选 质粒救援的探针和获得的突变基因的全长序列。 最后，将进行通过mRNA注射的表型响应分析 确认克隆基因的拯救能力。